Use of beta-1,3 (4)-endoglucanohydrolase, beta-1,3 (4)-glucan, diatomaceous earth, mineral clay and glucomannan to augment immune function

ABSTRACT

A method for the augmentation of immune function is described. The invention comprises a combination of β-1,3 (4)-endoglucanohydrolase, β-1,3 (4)-glucan, diatomaceous earth, mineral clay and glucomannan, which is fed to or consumed by mammalian or avian species in amounts sufficient to augment immune function. The invention described may be admixed with feeds or foods, incorporated into pelleted feeds or foods or administered orally to mammalian and avian species.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/566,433, filed Aug. 3, 2012, which is a continuation U.S. applicationSer. No. 11/668,375, filed Jan. 29, 2007, now U.S. Pat. No. 8,236,303,which is a divisional of U.S. application Ser. No. 10/829,633, filedApr. 5, 2004.

FIELD

This disclosure relates to methods and compositions for the augmentationof immune function in mammalian and avian species.

BACKGROUND

The immune system consists of two general features. These are: 1) theinnate immune system and 2) the adaptive (antibody-mediated) immunesystem. The innate system represents the first line of defense againstan invading pathogen (whether bacterial or fungal) and provides theadaptive immune system with enough time (3-5 days) for it to build upantibodies which are used to “fight” pathogens. While the innate andadaptive systems are often described separately, they function intandem; striving to sequester and neutralize a pathogen challenge.

The innate immune system. The innate immune system consists of severalinteresting components: Aspects include:

1. Physical and chemical barriers to pathogens provided by epithelium,gastric acid and digestive enzymes.

2. Cells which engulf and digest invading pathogens (e.g., neutrophils).

3. Receptors on the surface of these cells which recognize and bind topathogens.

4. Signaling molecules (e.g., chemokines, cytokines) which communicatesites of infection and regulate expression of immune genes.

Neutrophils. Neutrophils are among the most important cells of theinnate immune system. They are the first cell to arrive at a site ofinfection. In mammals, there are billions of neutrophils of which aboutone-half are freely circulating in the blood (Burton and Erskine, 2003).The remainder are held in reserve in bone marrow where they are formed.Neutrophils express an extracellular binding protein on their membranestermed “L-selectin” (also termed CD62L). The role of L-selectin is tointeract weakly with the endothelial cell wall thereby allowing theneutrophil to “roll” along the wall of a blood vessel and to “monitor”the cell wall for the presence of signals which indicate a localinfection (FIG. 1). The presence of pathogens in peripheral tissuescauses release of local chemicals which then signal a rolling neutrophilof an infection. In response to these signals, L-selectin is shed fromthe surface of the neutrophil (see FIG. 1) and other more adhesivemolecules are expressed on its surface. These molecules essentially“glue” the neutrophil within the blood vessel adjacent to the site ofinfection. The activated neutrophil then migrates through theendothelial cell wall toward the invading pathogen. Interleukin-1β isproduced by the neutrophil as a pro-inflammatory cytokine. This aids inmediating inflammation and in facilitating containment of invadingpathogens. During neutrophil migration, chemical signals originatingfrom the site of infection (such as TNF-β and interferon γ) activate theneutrophil to become a mature “killer cell”. The mature neutrophilmigrates toward the site of infection where it interacts withpathogen-associated microbial patterns (PAMPs) on the surface ofpathogens via several types of receptors. These receptors are expressedon the surface of the neutrophil and include the followingwell-identified types (FIG. 2):

a—CD18 and CD14

b—Toll-like receptors (TLRs)

c—C3b and C3bi (complement factors)

d—Fc

Binding of neutrophils to pathogens via receptors. Both CD14 and CD18receptors bind with lipopolysaccharide (LPS), a common polysaccharidestructure associated with membranes of gram-negative bacteria. Inaddition, neutrophils express toll-like receptors (TLRs) which recognizeand bind to additional structures associated with pathogens. So far, tendifferent toll-like receptors have been identified in mammals (FIG. 2and Table 1). TLRs play a critical role in early innate immunity toinvading pathogens by sensing microorganisms. These evolutionarilyconserved receptors recognize highly conserved structural motifs onlyexpressed by microbial pathogens, called pathogen-associated microbialpatterns (PAMPs: Invivogen, 2004). Stimulation of TLRs by PAMPsinitiates a signaling cascade that involves a number of proteins, suchas MyD88 and IRAK (FIG. 2). This signaling cascade leads to theactivation of the transcription factor NF-kB which induces the secretionof cytokines that direct the adaptive (i.e., antibody-mediated) immuneresponse. TLRs are predominantly expressed in tissues involved in immunefunction, such as spleen and peripheral blood leukocytes, as well asthose exposed to the external environment such as lung and thegastrointestinal tract. Ten human and nine mouse TLRs have beencharacterized, seven of which have had their ligands identified. Forexample, TLR2 is essential for the recognition of a variety of PAMPs,including bacterial lipoproteins, peptidoglycan, and lipotechoic acids.TLR3 is implicated in virus-derived double-stranded RNA. TLR4 ispredominantly activated by lipopolysaccharide. TLR5 detects bacterialflagellin and TLR9 is required for response to unmethylated CpG DNA(Table 1). Recently, TLR7 and TLR8 were shown to recognize syntheticantiviral molecules. These receptors are essential elements in hostdefense against pathogens by activating the innate immunity (Invivogen,2004).

Bovine TLRs. Relatively few studies on PAMPs have been completed withbovine cells. So far, bovine immune cells have been reported to containTLR2 and TLR4 (Werling et al., 2004). Polymorphisms have been reportedin bovine TLR4 which may determine susceptibility to bovine respiratorydisease and Johne's disease (White et al., 2003).

C3b and C3bi are components of the complement cascade whereas the Fcreceptor binds to the “constant region” of antibodies. Hence, pathogenswhich are coated with complement factors or antibody (i.e., pathogenswhich are opsonized) are also recognized by activated neutrophils andare subsequently phagocytosed. In other words, activated neutrophilspossess several means by which they recognize pathogens (Table 1).

Phagocytosis and killing. The binding of neutrophils, (and otherphagocytic cells) to cell-surface markers of pathogens via thesereceptors, then permits the phagocytic cell to engulf the invadingpathogen and “kill” it (FIG. 3). Presently, two mechanisms for “killing”are known. These include: 1) an oxidative burst, where the phagocyteexpresses reactive oxygen species which destroy the phagocytosedpathogen, and 2) fusion of the engulfed pathogen with a lysosome-likestructure to form a “phagosome”. The phagosome is rich in digestiveenzymes which mediate complete digestion of pathogens.

Common infections. Mammalian and avian species are continuallychallenged by pathogens in the gastrointestinal track and in the lung.These are important sites for resident neutrophils where minimizepathogen invasion. In addition, the mammary gland of mammals representsa site for pathogen challenge. In all infections, the innate immunesystem plays a key initial role in fighting-off the initial immunechallenge. The innate system is essential to allow the adaptive(antibody-mediated) system to develop and mount a more-specific anddirected immune response.

Cooperation between the innate and acquired immune system in ruminants.Antibodies which are specific to an invading pathogen leak into a siteof infection to optimize clearance of a pathogen. Individuals with ahigh titer against a specific antigen are able to deliver theseantibodies into the site of infection via a leaky endothelium (arisingfrom an inflammatory response). Arrival of reactive antibodies (i.e.,IgG2) in the alveolus coats (opsonizes) the pathogen and, as notedpreviously, allows neutrophil recognition of pathogens via Fc receptors(Table 1) and phagocytosis.

Stress and immune function. Stress reduces individuals' abilities tofight disease. The negative effects of stress on the immune system aremediate by the steroidal stress hormones (Cortisol, hydrocortisone andcorticosterone). Burton and co-workers at Michigan State University(Weber et al., 2001) have identified the mechanism by which stressbrings about a reduction in immune function. Specifically, they havedocumented that glucocorticoids (i.e., cortisol) “spike” nearparturition (FIG. 4) and reduce L-selectin expression in neutrophils(FIG. 5). This compromises one important aspect of an individual's firstline-of-defense against pathogen challenge. Specifically, a stressed,immunosuppressed individual has reduced ability to monitor endothelialcell lining for sites of infection and to attack and to sequesterpathogens. This may result in an infection (FIG. 6).

SUMMARY

The object of the present invention is to provide a novel and previouslyunknown method for augmentation of the immune system in mammalian andavian species. The invention may be applied to, but not limited to,mammalian and avian species and will reduce susceptibility of anindividual to both fungal and bacterial diseases.

A further object of this invention is to provide a method foraugmentation of immune function and to thereby minimize or obviatemorbidities and mortalities caused by, but not limited to, pathogenicfungi and bacteria with a preparation comprising a combination of β-1,3(4)-endoglucanohydrolase, β-glucan, diatomaceous earth, glucomannan, andmineral clay, such as aluminum silicate, montmorillonite clay, bentoniteor zeolite.

Another object of the invention is to provide a composition comprising acombination of β-1,3 (4)-endoglucanohydrolase, β-glucan, diatomaceousearth, mineral clay, and glucomannan, which additively augments immunefunction and thereby, reduces potential of pathogenic fungi and bacteriato cause morbidities and mortalities in mammalian and avian species.

Additional objects, advantages and novel features of the invention willbe set forth, in part, in the description that follows and will, inpart, become apparent to those skilled in the art upon examination ofthe following or may be learned with the practice of the invention. Toachieve the foregoing and other objects, and in accordance with thepurposes of the present invention as described herein, a novel method isdescribed for the augmentation of immune function of mammalian and avianspecies. In particular, this invention increases expression ofneutrophil L-selectin and interleukin-1β and thereby minimizes oreliminates the colonization of the epithelial surfaces and underlyingparenchymal tissues by pathogenic fungi and bacteria, reduces thepopulations of pathogenic organisms in blood and thereby minimizes oreliminates pathologies directly caused by and indirectly caused by thiscolonization. The invention comprises a mixture of β-1,3(4)-endoglucanohydrolase, β-glucan, diatomaceous earth, mineral clay,and glucomannan. The diatomaceous earth is standard commercial gradeavailable from a variety of sources. The β-1,3 (4)-endoglucanohydrolaseis produced from submerged fermentation of a strain of Trichodermalongibrachiatum. The β-1,3 (4)-glucan and glucomannan are derived from acommercial product and are an extraction from any of a number of yeastorganisms. The mineral clay product is a standard commercial grade(examples include, but are not limited to, montmorillonite clay,bentonite and zeolite). Extractions and productions of diatomaceousearth, yeast cell wall extract and mineral clay are well known in theart and commercially-available.

The compositions which are provided by the invention can be fed to anymammalian or avian species including, but not limited to, bovine,equine, ovine, caprine and avian species. When admixed with the feed orfood or fed as a supplement, the invention augments immune functionthereby reducing colonization by pathogens. The invention also minimizesor eliminates invasion of the blood compartment by pathogenic fungi andbacteria. The invention thereby minimizes or eliminates themanifestations of the pathologies typically associated with epithelialand systemic fungal and bacterial infections. Administration of theproduct may be used as a prophylactic (i.e., to prevent colonization andgrowth of pathogenic fungal and bacterial species in mammalian or avianspecies), as an additive to feeds or foods infected with pathogenicfungi or bacteria or as a preferred method to treat and thereby minimizeor eliminate an existing, diagnosed or non-diagnosed, fungal orbacterial infection. Application of the invention as described hereinand via the specific and novel mechanisms described herein will minimizeand possibly eliminate manifestations of fungal and bacterialinfections. Application of the invention as described herein will alsominimize or possibly eliminate manifestations associated with thepresence of pathogenic fungal and bacterial organisms, as identifiedabove, in food or feed of mammalian and avian species.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings and photographs which are incorporated intothe following “Detailed Description of the Invention” form part of thespecification and illustrate several aspects of the present inventionand, together with the Detailed Description, serve to explain thedetails of the invention.

FIG. 1 is a schematic drawing illustrating the movement of neutrophilsthrough a blood vessel. L-selectin (CD62L) is shown as circles on thesurface of the neutrophils A-E. These allow docking of the neutrophilswith endothelium. There is shedding of L-selectin and migration ofneutrophil into peripheral tissue toward a site of infection F. (Source:Burton and Erskine, 2003.)

FIG. 2 is a schematic diagram illustrating toll-like receptors (TLRs) onthe surface of an immune cell, and signal transduction following bindingof TLRs with microbial PAMPs (pathogen-associated molecular patterns).(Source: M. Adib-Conquy, C. Fitting, 2002.)

FIG. 3 illustrates a macrophage cell engulfing a bacterium in a processcalled phagocytosis. Toll-like and other receptor direct phagocytes torecognize microbes. Pseudopodial projections surround the bacterium.(Source: Travis, 2002.)

FIG. 4 is a bar graph of cortisol levels in dairy cattle relative to dayof parturition. Cortisol peaks at day of parturition. (Source: Weber etal., 2001.)

FIG. 5 is a bar graph of neutrophil surface CD62L (L-selectin)expression in cattle relative to day of parturition. (Source: Weber etal., 2001.)

FIG. 6 is a schematic diagram illustrating neutrophils lackingL-selectin expression in a stressed dairy cow. (Source: Burton andErskine, 2003.

FIG. 7. Effect of five experimental treatments on concentrations ofneutrophil L-selectin. An experiment was conducted with 60 sheep. Twelvesheep were allocated to each treatment. The treatments consisted of:

1. Control

2. Immunosuppressed (daily injections of Azium [Dexamethasone], 0.1mg/kg twice/day).

3. Immunosuppressed plus experimental product fed at 0.5% of daily drymatter intake.

4. Immunosuppressed plus moldy feed (addition of Aspergillusfumigatus-infected wheat mill run; 1.5 lbs/head/day).

5. Immunosuppressed plus moldy feed (as in Treatment 4) plus theexperimental feed product as outlined in Treatment 3.

The duration of the trial was 28-days. On Day 28, blood was taken fromsix sheep per treatment and neutrophils were recovered by Percollgradient centrifugation. The concentrations of L-selectin weredetermined by Western blot analysis using an antibody specific toL-selectin. Relative concentrations of L-selectin among the fivetreatment groups are shown in FIG. 8.

FIG. 8. Scanning densitometry of data shown in FIG. 7.

FIG. 9. Analysis of neutrophil interleukin-1β in the same sheepneutrophil samples presented in FIG. 7.

FIG. 10. Scanning densitometry of data shown in FIG. 9.

FIG. 11. Concentrations of Aspergillus fumigatus in blood samples takenfrom sheep on Day 28 of the above study. A. fumigatus DNA levels wereassessed using a quantitative Sybr-Green PCR-based assay specific for A.fumigatus.

DETAILED DESCRIPTION

The present invention is based on the novel discovery that a combinationof β-1,3 (4)-endoglucanohydrolase, β-1,3 (4)-glucan, diatomaceous earth,mineral clay, and glucomannan effectively augments immune function andreduces colonization of tissues and blood by a pathogen.

The β-1,3 (4)-endoglucanohydrolase is from a commercial source and isproduced from submerged fermentation of a strain of Trichodermalongibrachiatum.

The diatomaceous earth is prepared by methods commonly known in the art.It is available as a commercially-available acid-washed, product with95% silica (SiO₂) and with its remaining components not assayed butconsisting primarily of ash (minerals) as defined by the Association ofAnalytical Chemists (AOAC, 2002).

The yeast cell wall extract is prepared by a method commonly known inthe art. It is a commercial source of β-1,3 (4) glucan and glucomannanderived from primary inactivated yeast (Saccharomyces cerevisiae) withthe following chemical composition:

Moisture 2-3%

Dry matter 97-98%

Proteins 14-17%

Fats 20-22%

Phosphorus 1-2%

Mannans 22-24%

β-(4) glucan 24-26%

Ash 3-5%

The mineral clays (aluminosilicates) used in this invention may befulfilled by any of a variety of commercially-available clays including,but not limited to, montmorillonite clay, bentonite and zeolite.

In a preferred embodiment of the invention, β-1,3(4)-endoglucanohydrolase, diatomaceous earth, yeast cell wall extractand mineral clay are combined at 0.05-3%, 1-40%, 1-20% and 40-92%,respectively. In a preferred composition, β-1,3(4)-endoglucanohydrolase, diatomaceous earth, yeast cell wall extractand mineral clay are combined at 0.1-3%, 5-40%, 2-10% and 40-80%,respectively. In an especially preferred embodiment of the invention,β-1,3 (4)-endoglucanohydrolase, diatomaceous earth, yeast cell wallextract and mineral clay are combined at 0.2-3%, 30-40%, 4-6% and50-65%, respectively. The preferred physical form of the invention is adry, free-flowing powder which is suitable for direct inclusion into afeed, food product or as a supplement to a total mixed ration or diet.

The compositions provided by the present invention may be incorporateddirectly into commercially-available feeds or food products or fed assupplements to commercially-available feeds or food products. Thecomposition contained in the present invention may be fed to anymammalian or avian species. The methods of the invention compriseaugmenting immune function in mammalian and avian species. Whenincorporated directly into feeds, the present invention may be added tofeeds in amounts ranging from 0.1 to 5 kg per ton of feed. In anespecially preferred composition, the invention may be added to feeds inamounts ranging from 1-2 kg per ton of feed.

The composition contained in the present invention may be added toanimal feedstuffs or to foods in amounts ranging from 0.0125% to 2% byweight of feed. In a preferred embodiment, the composition is added toanimal feedstuffs or to food in amounts from 0.0625% to 1% by weight offeed. In an especially preferred embodiment, the invention is added inamounts from 0.125% to 0.5% by weight of feed.

Alternatively, the composition contained in the present invention may befed directly to mammalian or avian species as a supplement in amounts0.016 grams/kg to 0.37 grams/kg of live body weight per day. In anespecially preferred embodiment, the invention may be provided tomammalian and avian species in amounts of 0.10 grams/kg to 0.20 grams/kgof body weight per day. One of skill in the art can appreciate that theamount of the invention fed can vary depending upon the animal species,size of the animal and type of the feedstuff to which the invention isadded.

The novel methods of this invention comprise the ability of acombination of β-1,3 (4)-endoglucanohydrolase, diatomaceous earth, yeastcell wall extract and clay to augment immune function. The benefitsresulting from the application of the invention to mammalian speciesinclude, but are not limited to, reduced death losses, reduced incidenceof mycotic abortion, reduced incidence of jejunal hemorrhage syndrome(dead gut syndrome), reduced incidence of scouring (diarrhea), improvedgrowth rate, improved efficiency of growth, improved milk production,improved efficiency of milk production and reduced somatic cell countsin milk products (dairy animals). The benefits from the application ofthe invention to avian species include, but are not limited to, reduceddeath losses, improved growth and egg production, improved fertility,and reduced incidence of enteric diseases.

The following are intended to be illustrative of the invention, and arenot to be considered restrictive of the scope of the invention asotherwise described herein.

EXAMPLE 1

An experiment was conducted using 60 growing male and female sheep.Sheep were allocated to one of the five treatments (seven females andfive males per treatment):

1. Control.

2. Immunosuppressed (daily injections of Azium [Dexamethasone], 0.1mg/kg twice/day).

3. Immunosuppressed plus the invention fed at 0.5% of daily dry matterintake.

4. Immunosuppressed plus moldy feed (addition of Aspergillusfumigatus-infected wheat mill run; 1.5 lbs/head/day).

5. Immunosuppressed plus moldy feed (as in Treatment 4) plus theinvention as outlined in Treatment 3.

Animals were fed a dairy-type diet for a period of 28 days.Immunosuppression was mediated in Treatments 2, 3, 4 and 5 by dailyinjection of Azium using a high dose (a model of extreme stress: Weberet al., 2001). Sheep on Treatments 4 and 5 were challenged with apathogenic mold by feeding wheat mill run which had been contaminatedwith a pathogenic mold (Aspergillus fumigatus). Sheep on Treatments 3and 5 were supplemented with the invention at a rate of 0.5% of theirdaily dry matter intake. Following 28 days, blood samples were taken viajugular puncture and the neutrophil fractions were isolated usingPercoll density gradient centrifugation. Following this, samples ofneutrophil protein were processed using sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blotting using antibodieswhich are specific for L-selectin and interleukin-1-β. Relativeconcentrations of L-selectin and interleukin-1-β. were assessed usingscanning densitometry.

FIGS. 7 and 8 demonstrate effects of the five experimental treatments onneutrophil L-selectin. Injection with Azium caused a marked reduction(P<0.05) in L-selectin and provides evidence that Azium injection was,in fact, immunosuppressive. Addition of mold to the diets had no effect(P>0.05) on L-selectin concentrations. Of interest, addition of theinvention to feed (Treatments 3 and 5) caused restoration (augmentation:P<0.05) of L-selectin.

Interpretation: The novel invention successfully restored (augmented)normal levels neutrophil L-selectin. Restoration of L-selectin onneutrophil surfaces will re-establish their ability to monitor theendothelial lining for pathogens.

FIGS. 9 and 10 demonstrate effects of the five experimental treatmentson neutrophil interleukin-1-β concentrations. Azium treatment caused amarked reduction (P<0.05) in neutrophil interleukin-1-β concentration.This demonstrates that Azium was immunosuppressive. The novel inventionhad no effect (P>0.05) on neutrophil interleukin-1-β in the absence of apathogen challenge (i.e., Treatment 3 versus Treatment 2); however, theinvention caused a marked increase (P<0.05) in neutrophilinterleukin-1-β in the presence of a pathogen challenge (i.e., Treatment5 versus Treatment 4).

Interpretation. Interleukin-1β is an important pro-inflammatory cytokinewhich enables the neutrophil to fulfill its role as a phagocyte. Abilityof the feed product to restore interleukin-1β in the presence of apathogen (A. fumigatus) demonstrates that pathogens potential effects ofthe invention on immune function.

FIG. 11 shows the effects of the five experimental treatments on bloodconcentrations of A. fumigatus. A. fumigatus concentrations weredetermined using a Sybr-Green real-time quantitative polymerase chainreaction (PCR) assay developed in our laboratory. The resultsdemonstrate that the invention reduced (P<0.05) A. fumigatusconcentration in blood.

Interpretation. The restoration of neutrophil function shown in FIGS.7-10 manifests itself by reducing pathogen load detected within theblood compartment. The invention reduces pathogen load.

These results show that the composition of the invention (i.e., mineralclay, yeast cell wall extract, diatomaceous earth and β-1,3(4)-endoglucanohydrolase) is capable of a previously-undescribed effectof augmenting immune function. The invention specifically restoreslevels of L-selectin and interleukin-1-β in neutrophils therebyrestoring the ability of neutrophils to monitor for the presence ofinvading pathogens.

The combination of products augments immunity in mammalian and domesticspecies and thereby prevents the invasion and colonization of the bloodcompartment. It represents a mixture which is flowable and easilyincorporated into feed products and food products. The present inventionwas effective in achieving its immunostimulatory effects under growthconditions which might be found in mammalian and avian digestive systemswhere nutrients, moisture, oxygen and elevated temperatures are providedby the host.

The foregoing description of the preferred embodiment of the inventionhas been presented for purposes of illustration and description. It isnot intended to be exhaustive or to limit the invention to the preciseform disclosed. Obvious modifications or variations are possible inlight of the above illustrations. The embodiment was chosen anddescribed to provide the best illustration of the principles of theinvention and its practical application to thereby enable one ofordinary skill in the art to utilize the invention in variousembodiments and with modifications as are suited to the particular usecontemplated. All such modifications and variations are within the scopeof the invention as determined by the appended claims when interpretedin accordance with the breadth to which they are fairly, legally andequitably entitled.

What is claimed is:
 1. A method for reducing a stress effect in ananimal, comprising administering to an animal having an increased stressindicator a composition comprising β-glucans, β-1,3(4)-endoglucanohydrolase, silica, a mineral clay, and mannans, whereinthe animal is selected from mammalian and avian species, therebyreducing a stress effect in the animal.
 2. The method of claim 1 wherethe increased stress indicator is an elevated stress hormone level andthe stress effect is immunosuppression mediated by the elevated stresshormone level.
 3. The method of claim 2 where the elevated stresshormone is cortisol, hydrocortisone, corticosterone, or a combinationthereof.
 4. The method of claim 2 where the elevated stress hormonelevel includes elevated glucocorticoids.
 5. The method of claim 1 wherethe composition is administered to the animal at or around a time ofparturition.
 6. The method of claim 1, further comprising administeringthe composition to the animal as a prophylactic to prevent colonizationor growth of pathogenic fungal or bacterial species in the animal. 7.The method of claim 1 where administering the composition augments theanimal's innate immune function.
 8. The method of claim 7 whereaugmenting the animal's innate immune function includes increasing theexpression of L-selectin, interleukin-1β, or a combination thereof. 9.The method of claim 1 where the silica is provided by diatomaceousearth.
 10. The method of claim 1 where the mannans comprise glucomannan.11. The method of claim 1 where the mineral clay comprisesmontmorillonite, bentonite, aluminosilicate, or zeolite clays, ormixtures thereof.
 12. The method of claim 1 where the compositioncomprises from about 15% to about 40% silica, from about 50% to about81% mineral clay, from about 1.0% to about 5.0% β-glucans, from about0.05% to about 3.0% β-1,3 (4)-endoglucanohydrolase, and from about 1% toabout 8.0% mannans.
 13. The method of claim 1 where administering thecomposition comprises admixing the composition into foods or animalfeedstuffs that are subsequently fed to the animal.
 14. The method ofclaim 15 where the composition is admixed into the foods or animalfeedstuffs in a concentration of from about 0.0125% to about 5% byweight of the foods or animal feedstuffs.
 15. The method of claim 1where administering the composition comprises feeding the composition tothe animal as a supplement in an amount from about 0.016 grams/kg toabout 0.37 grams/kg of live body weight per day.
 16. The method of claim1 where administering the composition comprises administering thecomposition to the animal in an amount from about 0.125% to about 0.5%by weight of feed.
 17. The method of claim 1 where the composition isadministered daily to the animal.
 18. The method of claim 1 where thestress effect is susceptibility to a fungal or bacterial disease or abacterial disease and administering the composition reduces the animal'ssusceptibility to the fungal or bacterial disease.
 19. The method ofclaim 1 where the animal is a bovine, equine, ovine, caprine, or avianspecies.
 20. A method for reducing a stress effect in an animal,comprising: identifying a stressed animal of a mammalian or avianspecies by determining an elevated stress hormone level; andadministering to the stressed animal a composition comprising β-glucans,β-1,3 (4)-endoglucanohydrolase, silica, a mineral clay, and mannans,thereby reducing a stress effect in the animal.